Objective:To analyze the mutation characteristics of rpoB gene in rifampicin-resistant Brucella strains. Methods:DNA of 4 rifampicin-resistant Brucella strains (JSY-26, G-9, WSY-13 and AW-3) isolated from Xinjiang Uygur Autonomous Region was selected, rifampicin rpoB gene was amplified by PCR and its nucleotide sequence was sequenced. The rpoB gene sequences of rifampicin-resistant Brucella standard strain (RB51) and sensitive strain (ALT-8) were used as reference, the mutation sites and types of the rpoB gene inside and outside the rifampicin resistance determination region (RRDR) of the 4 rifampicin-resistant Brucella strains were analyzed by Mega 7.0 software. Results:Through sequence alignment, both JSY-26 and WSY-13 strains underwent a single base point mutation at the RRDR 1 576 bp of the rpoB gene, with the base changing from guanine (G) to adenine (A). The G-9 strain underwent a single base point mutation at the RRDR 1 606 bp of the rpoB gene, with the base changing from cytosine (C) to A. The AW-3 strain showed 5 mutations of 3 types outside rpoB gene RRDR at 2 536, 2 537, 2 626, 2 636 and 2 654 bp, namely 3 insertion mutations [thymine (T) insertion once and C insertion twice], 1 deletion mutation (C deletion), and 1 single base point mutation (from G to C mutation).Conclusion:The RRDR mutations in the rpoB gene of the rifampicin-resistant Brucella strains are mainly characterized by single base point mutations, while multiple insertion and deletion mutations occur outside the RRDR.

Objective:To investigate the role of Pink1/Parkin-induced mitophagy in acute lung injury of exertional heat stroke rats.Methods:Sixty SD rats were randomly divided into four groups, including normal group (CON group), normal Parkin overexpression group (CON+Parkin group), heat stroke group (EHS group) and heat stroke Parkin overexpression group (EHS+Parkin group), with fifteen rats in each group. The rat model of exertional heat stroke was established and the survival curve was drawn. Pulmonary coefficient and pulmonary capillary permeability were detected. HE staining was used to observe the pathological changes of lung tissue. ELISA was used to detect the contents of IL-6, IL-1β, TNF-α and ROS in lung tissue; immunohistochemistry was used to observe apoptosis in lung tissue; Western blot was used to determine the expression of Pink1, Parkin, P62 and LC3 in rat lung tissue, and the LC3II/LC3I ratio was calculated. Single factor multi-level group comparison was performed by single factor analysis of variance, SNK-q method was used to further pairwise comparison between groups.Results:Compared with the normal group, the survival rate of EHS group was decreased ( P<0.001), lung coefficient and pulmonary vascular permeability were increased [(4.39±0.42), (33.38±8.29) μg/g, P<0.05)], lung tissue was exudative and solid, the levels of inflammatory factors IL-6, IL-1β, TNF-α and ROS were significantly increased[(34.31±5.34) pg/mL, (34.03±4.78) pg/mL, (91.64±8.16) pg/mL, (259.01±89.17) U/mg, P<0.05)], and apoptosis was increased. Western and immunohistochemistry results showed that the expressions of Pink1 and Parkin were decreased, co-location of Pink1and Parkin was attenuated, LC3II/LC3I were decreased, and P62 expression was increased. Compared with the EHS group, the survival rate of EHS+Parkin group was significantly increased ( P<0.05), lung coefficient and pulmonary vascular permeability were decreased [(3.83±0.62), (22.49±7.90) μg/g, P<0.05)], exudation and consolidation and other pathological changes were significantly reduced, and the levels of the above inflammatory factors and ROS were significantly decreased [(14.09±3.24) pg/mL, (26.94±2.11) pg/mL、(63.35±11.62) pg/mL, (161.13±26.31) U/mg, P<0.05]. Lung tissue apoptosis was reduced. The co-location of Pink1and Parkin、Parkin expression and LC3II/LC3I ratio were increased ( P<0.05), P62 expression was decreased( P<0.05), while Pink1 expression was not significant different (q=0.75). There was no difference between normal group and normal Parkin overexpression group (q=0.95). Conclusion:Activation of Pink1/Parkin-induced mitophagy can alleviate the acute lung injury in exertional heat stroke rats.

ObjectiveTo observe the effect of Sanhuang Tangshenkang on the Wnt/β-catenin signaling pathway in the bone tissue of diabetic rats. MethodA high-sugar and high-fat diet was administered for 4 weeks， along with intraperitoneal injection of freshly prepared 2% streptozotocin （pH 4.5） at 30 mg·kg^-1 body weight to induce a diabetes model in rats. The rats with diabetes were randomly divided into model group， low- and high-dose Sanhuang Tangshenkang groups （12.8， 38.4 g·kg^-1）， and Gushukang group （1.8 g·kg^-1） according to the blood glucose level. Rats of the same age were fed on a regular diet and assigned to the control group. After 12 weeks of respective treatments with drugs or physiological saline， the fasting blood glucose （FBG） levels of the rats were measured using an automated biochemical analyzer. Enzyme-linked immunosorbent assay （ELISA） was used to detect fasting serum insulin （FINS）， bone-specific alkaline phosphatase （BALP）， and tartrate-resistant acid phosphatase （TRAP） levels. Micro-computed tomography （Micro-CT） was used to scan the femurs of rats to observe bone tissue microstructure and measure bone mineral density （BMD）. Hematoxylin-eosin （HE） staining and safranin O/fast green staining were performed to observe pathological changes in the femoral bone tissue. Immunohistochemistry （IHC） and Western blot were used to detect the expression of Wnt3a， low-density lipoprotein receptor-related protein 5 （LRP-5）， and β-catenin proteins. ResultCompared with the control group， the model group showed a significant increase in FBG， FINS， and TRAP levels （P<0.01）， a significant decrease in BALP level （P<0.01）， a significant decrease in BMD （P<0.01）， and disorganized， elongated， and sparse bone trabecular structures with fractures and increased lipid droplets. Additionally， the expression of Wnt3a， LRP-5， and β-catenin proteins decreased （P<0.05， P<0.01）. Compared with the model group， the low- and high-dose Sanhuang Tangshenkang groups showed a reduction in FBG and an increase in BALP （P<0.05）. The low-dose Sanhuang Tangshenkang group also exhibited a decrease in FINS （P<0.05）. All treatment groups showed a significant decrease in TRAP （P<0.01）， varying degrees of improvement in BMD （P<0.05， P<0.01））， increased and denser bone trabeculae with more regular arrangements and reduced lipid droplets， and improved bone microstructure morphology. The average optical density values of Wnt3a， LRP-5， and β-catenin proteins were significantly increased in all drug-treated groups （P<0.05， P<0.01）， and the expression of Wnt3a， LRP-5， and β-catenin proteins was elevated （P<0.05， P<0.01）. ConclusionSanhuang Tangshenkang may regulate the imbalance of the Wnt/β-catenin signaling pathway by increasing the expression of Wnt3a， LRP-5， and β-catenin proteins in bone tissue， which may promote bone formation， reduce bone resorption， and lower blood glucose levels， thereby achieving the effect of preventing and treating diabetic osteoporosis.

ObjectiveTo establish a set of single nucleotide polymorphisms (SNP) detection protocol for inbred rats based on multiplex PCR-ligase detection reaction (LDR). MethodsA total of 40 rats SNP sites were selected on chromosomes 1-20 and X of rats among 5 inbred strains of rats, and the 40 SNP sites were randomly divided into four groups. A genetic detection protocol for 4 groups of SNP in inbred rats based on multiplex PCR-LDR technology was constructed. 9 commonly used rat strains from two other domestic rat suppliers were detected by this protocol. Finally, the feasibility of this protocol was verified by comparing the amplification effects of different DNA polymerases by a third-party laboratory. ResultsWhen using the constructed SNP detection protocol for inbred rats to test 5 rat strains, all sites in each sample obtained good amplification results. The 9 commonly used rat strains from two other rat suppliers in china were also well amplified by this SNP detection protocol, and 40 SNPs were homozygous in each Inbred strain. The results of detection of the same rat DNA samples with three different DNA polymerases showed that the Multiplex PCR Kit, AmpliTaq Gold 360 DNA polymerase and Platinum II Taq hot start DNA polymerase had electrophoretic peaks of amplification products at all SNP sites in groups 1 to 3, and Platinum II Taq hot start DNA polymerase had one less electrophoretic peak of the amplification products at the SNP sites in group 4. In addition, inter-laboratory comparisons showed consistent results for the same amplification system. ConclusionBased on multiplex PCR-LDR technology, this study successfully established a SNP detection protocol for rats covering all autosomes and X chromosomes with the excellent stability and repeatability.

Objective:To investigate the epidemic status and molecular characteristics of Borrelia burgdorferi in ticks in Xinjiang Uygur Autonomous Region (referred to as Xinjiang). Methods:From April to June 2020, 312 samples of Ixodes were collected in 6 areas of Yili, Alashankou, Hutubi, Qinghe, Fuhai and Wujiaqu, Xinjiang. Nested PCR and fluorescence quantitative PCR were used to detect Borrelia burgdorferi in ticks. The positive samples by both methods were genotyped and identified by nested PCR products. Results:The positive rates of nested PCR and fluorescence quantitative PCR were 8.97% (28/312) and 11.86% (37/312), respectively. Among them, the fluorescence quantitative PCR positive rate of Qinghe was the highest of 35.29% (12/34), and the positive rate of Fuhai was the lowest of 2.00% (1/50). The positive samples by both methods was 26. Genotyping results showed that 12 samples were highly homologous to Borrelia garinii, 10 copies to Borrelia burgdorferi sensu stricto, and 4 copies to Borrelia afzelii. Conclusions:The positive rate of Borrelia burgdorferi in ticks in Xinjiang is higher, which has confirmed that there are 3 pathogenic Borrelia burgdorferi genotypes in Xinjiang. The dominant genotype is Borrelia garinii, followed by Borrelia burgdorgferi sensu stricto genotype and Borrelia afzelii genotype.

Lactic acid bacteria (LAB) are a representative probiotic. As the dominant flora in the human intestinal tract, LAB can regulate the balance of human intestinal flora and improve host health. The purpose of this study was to isolate and screen LAB that are well suited to the intestinal characteristics of the Chinese population, with excellent probiotics and high antibacterial activity. After 16S ribosomal RNA (rRNA) homology and phylogenetic tree analysis, potential probiotics were tested for their antibacterial activity, resistance to artificial gastrointestinal fluid and drugs, surface hydrophobicity, and safety. Three strains of LAB with acid resistance, bile salt resistance, epithelial cell adhesion, and no multidrug resistance were selected:

Objective : To evaluate the clinical effects of resin infiltration in treating molar-incisor hypomineralization. Methods : Twelve patients (36 teeth) with mild molar-incisor mineralization imperfecta who met the inclusion criteria were selected and treated with penetrating resin. Before treatment (T0), at the one-week follow-up (T1) and at the six-month follow-up (T2), a Crystaleye spectrophotometer was used to take photos, and the color differences (ΔE) between the lesion area and the surrounding normal enamel area at different times were calculated and analyzed. The area of the lesion and the total area of the labial surface of affected teeth were calculated using Adobe Photoshop CS3 software. The corresponding area ratio (R) and treatment efficiency (SR) were obtained. The R value was used to judge the effectiveness of resin penetration in the treatment of molar and incisor mineralization imperfecta and to evaluate its aesthetic effect. The results were analyzed by repeated measures one-way ANOVA. Results: The color difference of the lesion area vs sound adjacent enamel (ΔE) decreased significantly, and that of the lesion area decreased significantly after resin infiltration (P < 0.001). The success rate was approximately 86.02%, which means that the esthetic effect of resin infiltration in treating molar-incisor hypomineralization was remarkable. No important adverse events or side effects were observed. Conclusion The aesthetic effect of resin infiltration in the treatment of mild molar and incisor hypomineralization is good. This method is recommended for clinical use.

Objective:The epidemiological characteristics and drug susceptibility changes of Pseudomonas aeruginosa(PA) infection in the respiratory tract of children in suzhou were analyzed in order to provide guidance and suggestions for the rational use of antibiotics in this region. Methods:Bacterial culture results of nasopharyngeal secretions from 21 176 children admitted to the Department of Respiratory, Children′s Hospital Affiliated to Soochow University for respiratory tract infections from January 2008 to December 2017 were collected.According to age, season, underlying disease conditions and the presence of intensive care units (ICU) during the hospitalization, these children were divided into different groups.The epidemic characteristics of PA infection were monitored, and changes in PA drug sensitivity were dynamically observed.Results:Among 21 176 nasopharyngeal secretions from children with respiratory tract infection, 191 cultures were detected with PA, and showing a positive detection rate of 0.90% (191/21 176 cases). The annual detection rate was different.The highest detection rate was 2.24% (50/2 234 cases) in 2009 and the lowest was 0.41% (9/2 207 cases) in 2014.The detection rate of PA in children with in >6 months to 1-year-old group was the highest (1.52%, 53/3 497 cases), and the lowest was 0.57%(11/1 934 cases) in the >5-year-old group.The detection rates of PA in spring, summer, autumn, and winter were 1.11% (60/5 420 cases), 1.21% (61/5 046 cases), 0.81% (46/5 670 cases), and 0.48% (24/5 040 cases), respectively.The difference of PA detection rates between summer (the highest rate) and the winter (the lowest rate) was statistically significant ( χ2=18.611, P<0.001). Among the 21 176 children, 18.89% (4 000/21 176 cases) had basic diseases, and the PA detection rate in such kind of patients was 1.28% (51/4 000 cases), which was higher than that in patients without basic diseases (0.82%, 140/17 176 cases). The PA detection rate in the ICU group was 4.41% (15/340 cases), which was significantly higher than that in the general group (0.84%, 176/20 836 cases) ( χ2=7.678, 47.623, all P<0.05). There were no strains susceptible to Ampicillin, Ampicillin/Sulbactam and compound Sulfamethoxazole, and no strains susceptible to Ceftriaxone from 2010 to 2017.The susceptibility rate to Imipenem was low from 2012 to 2015, and the lowest was only 66.7% in 2014.The sensitivity rate to Aztreonam fluctuated significantly from year to year.Strains were all highly sensitive to Piperacillin, Piperacillin Tazobactam, Ciprofloxacin, Levofloxacin, Gentamicin, and Tobramycin. Conclusions:PA infection is prone to occur in young children with basic diseases and relatively severe illness.Summer is a high-incidence season.PA is generally highly resistant to commonly used antibiotics.

Objective To identify molecular typing of Brucella abortus isolates in Xinjiang,and determine the identification ability of multiple locus variable-number tandem repeat analysis (MLVA).Methods The optimized Brucella AMOS-PCR was used for identification of Brucella (n =7) genus and species in Xinjiang from 2010-2015,and MLVA-16 was used to further identify the isolates.Results were compared with the data of the Brucella standard strain provided by the http://mlva.u-psud.fr database.Cluster analysis was carried out with Bionumerics 6.6.Results The results of AMOS-PCR and MLVA-16 were identical,all were Brucella abortus.Further classification results of the MLVA-16 showed that the strain in Xinjiang was type 3 of Brucella abortus,which was basically the same as that of the domestic Brucella.Conclusions The molecular typing of isolates separated in Xinjiang is type 3 of Brucella abortus.MLVA can identify Brucella at the level of species,and highly sensitive to Brucella biotype and isolates differences,which provides a basis for the traceability and evolution of brucellosis epidemic strains.

Objective To explore the drug sensibility of Brucella from bovine and sheep in Xinjiang.Methods Using paper diffusion method,19 drugs of 8 kinds of antibiotics including aminoglycosides,macrolides,sulfonamides,tetracyclines,β-lactams,fluoroquinolones,chloramphenicols and rifamycins,were tested.Drug sensitivity test was conducted on 57 Brucella strains isolated from bovine and sheep in Xinjiang from 2010 to 2016.Results The 57 Brucella strains were highly sensitive to doxycycline,tetracycline,streptomycin,tobramycin,gentamicin,amikacin,amoxicillin,ofloxacin,fleroxacin,ciprofloxacin and chloramphenicol,with the sensitivity rates were all higher than 90％;and they were highly resistance to azithromycin,clarithromycin and bactrim,with the drug resistance rates were all higher than 80％.Conclusion Brucella from bovine and sheep in Xinjiang is sensitive to tetracyclines,aminoglycosides,β-1actams,fluoroquinolones and chloramphenicols.